Search results for: in vivo metabolites

Authors: Daphne Meza, Louie Abejar, David A. Rubenstein, Wei Yin

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Endothelial cell (ECs) morphology and function is highly impacted by the mechanical stresses these cells experience in vivo. Any change in the mechanical environment can trigger pathological EC responses. A detailed understanding of EC morphological response and function upon subjection to individual and simultaneous mechanical stimuli is needed for advancement in mechanobiology and preventive medicine. To investigate this, a programmable device capable of simultaneously applying physiological fluid shear stress (FSS) and cyclic strain (CS) has been developed, characterized and validated. Its validation was performed both experimentally, through tracer tracking, and theoretically, through the use of a computational fluid dynamics model. The effectiveness of the device was evaluated through EC morphology changes under mechanical loading conditions. Changes in cell morphology were evaluated through: cell and nucleus elongation, cell alignment and junctional actin production. The results demonstrated that the combined FSS-CS stimulation induced visible changes in EC morphology. Upon simultaneous fluid shear stress and biaxial tensile strain stimulation, cells were elongated and generally aligned with the flow direction, with stress fibers highlighted along the cell junctions. The concurrent stimulation from shear stress and biaxial cyclic stretch led to a significant increase in cell elongation compared to untreated cells. This, however, was significantly lower than that induced by shear stress alone, indicating that the biaxial tensile strain may counteract the elongating effect of shear stress to maintain the shape of ECs. A similar trend was seen in alignment, where the alignment induced by the concurrent application of shear stress and cyclic stretch fell in between that induced by shear stress and tensile stretch alone, indicating the opposite role shear stress and tensile strain may play in cell alignment. Junctional actin accumulation was increased upon shear stress alone or simultaneously with tensile stretch. Tensile stretch alone did not change junctional actin accumulation, indicating the dominant role of shear stress in damaging EC junctions. These results demonstrate that the shearing-stretching device is capable of applying well characterized dynamic shear stress and tensile strain to cultured ECs. Using this device, EC response to altered mechanical environment in vivo can be characterized in vitro.

Keywords: cyclic stretch, endothelial cells, fluid shear stress, vascular biology

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Authors: Chandrima Das, Sirshendu Chatterjee

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Tea (Camellia sinensis) is known as nature's low calorie wonder drink. Since ancient times hot consumptions of tea is very much popular. We have observed that many heat sensitive secondary metabolites which get destroyed on heating, moreover by people, who are permanently live at higher altitude or the members of high altitude expedition team, are deprived of various tea brewing facilities like electricity, fuel, etc. and the hence cold decoction of tea might be a good alternative. In this backdrop present study aims at the analysis of antioxidants like polyphenols, flavonoids and free radical scavenging activity as well as the l-theanine concentration of different types of cold brewed teas like black, green, white and oolong and compared with its hot decoction. Further, we also analysed in details about the bioactive components by using HPLC followed by green synthesis of nanoparticles. The study highlighted that the difference between the concentration of antioxidant in cold and hot brewed tea is insignificant and hence intake of cold decoction will be beneficial to health.

Keywords: antioxidants, flavanoid, polyphenols, HPLC, nanoparticles

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Authors: Gabrielle Robertson, Matthew Downs, Joseph Dagher

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Iron deposition in the brain has been linked with a host of neurological disorders such as Alzheimer’s, Parkinson’s, and Multiple Sclerosis. While some treatment options exist, there are no objective measurement tools that allow for the monitoring of iron levels in the brain in vivo. An emerging Magnetic Resonance Imaging (MRI) method has been recently proposed to deduce iron concentration through quantitative measurement of magnetic susceptibility. This is a multi-step process that involves repeated modeling of physical processes via approximate numerical solutions. For example, the last two steps of this Quantitative Susceptibility Mapping (QSM) method involve I) mapping magnetic field into magnetic susceptibility and II) mapping magnetic susceptibility into iron concentration. Process I involves solving an ill-posed inverse problem by using regularization via injection of prior belief. The end result from Process II highly depends on the model used to describe the molecular content of each voxel (type of iron, water fraction, etc.) Due to these factors, the accuracy and repeatability of QSM have been an active area of research in the MRI and medical imaging community. This work aims to estimate iron concentration in the brain via a single step. A synthetic numerical model of the human head was created by automatically and manually segmenting the human head on a high-resolution grid (640x640x640, 0.4mm³) yielding detailed structures such as microvasculature and subcortical regions as well as bone, soft tissue, Cerebral Spinal Fluid, sinuses, arteries, and eyes. Each segmented region was then assigned tissue properties such as relaxation rates, proton density, electromagnetic tissue properties and iron concentration. These tissue property values were randomly selected from a Probability Distribution Function derived from a thorough literature review. In addition to having unique tissue property values, different synthetic head realizations also possess unique structural geometry created by morphing the boundary regions of different areas within normal physical constraints. This model of the human brain is then used to create synthetic MRI measurements. This is repeated thousands of times, for different head shapes, volume, tissue properties and noise realizations. Collectively, this constitutes a training-set that is similar to in vivo data, but larger than datasets available from clinical measurements. This 3D convolutional U-Net neural network architecture was used to train data-driven Deep Learning models to solve for iron concentrations from raw MRI measurements. The performance was then tested on both synthetic data not used in training as well as real in vivo data. Results showed that the model trained on synthetic MRI measurements is able to directly learn iron concentrations in areas of interest more effectively than other existing QSM reconstruction methods. For comparison, models trained on random geometric shapes (as proposed in the Deep QSM method) are less effective than models trained on realistic synthetic head models. Such an accurate method for the quantitative measurement of iron deposits in the brain would be of important value in clinical studies aiming to understand the role of iron in neurological disease.

Keywords: magnetic resonance imaging, MRI, iron deposition, machine learning, quantitative susceptibility mapping

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Authors: M. Eickermann, M. K. Class, J. Junk

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Winter oilseed rape, Brassica napus L., is characterized by a high number of herbicide applications. Therefore, its cultivation can lead to massive contamination of ground water and soil by herbicide and their metabolites. A multi-side long-term field experiment (EFFO, Efficient crop rotation) was set-up in Luxembourg to quantify these effects. Based on soil sampling and laboratory analysis, preliminary results showed reduced dehydrogenase activities of several soil organisms due to herbicide treatments. This effect is highly depending on the soil type. Relation between the dehydrogenase activity and the amount of microbial carbon showed higher variability on the test side with loamy Brown Earth, based on Bunter than on those with sandy-loamy Brown Earth, based on calciferous Sandstone.

Keywords: cropping system, dehydrogenase activity, herbicides, mechanical weed control, oilseed rape

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Authors: Adnan Bekhit, Eskedar Lodebo, Ariaya Hymete, Hanan Ragab, Alaa El-Din Bekhit

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Malaria and leishmaniasis have emerged as serious universal health problems throughout history of mankind. According to the WHO 2008 malarial report, half of the world population is at risk of malarial infection with an estimate of 1 million deaths occurring annually mainly in the African region. Furthermore, 12-15 million people are infected with Leishmaniasis worldwide. Despite the continuous introduction of a large number of agents for the treatment of malaria, there is still unmet medical needs due to the emergence of resistance. Resistance has occurred for almost all therapeutic agents approved for the treatment of malaria. Accordingly, it was the aim of this work to design and synthesis a group of antimalarial-antileshmanial agents that would show inhibitory activity against chloroquine-resistant strain of Plasmodium falciparum. The synthesized compounds were designed to contain a pyrazolylpyrazoline moiety having an aromatic group (p-tolyl or p-chlorophenyl) at N1-position of one pyrazoline ring due to the reports of promising activities of such compounds. A formyl or acyl substituent was introduced at the N1-position of the other pyrazoline ring, to investigate the effect of bulkiness of acyl substituents at this position. The synthesized compounds were evaluated for their in-vivo antimalarial activity against Plasmodium berghei infected mice at dose levels of 20 and 30 mg/Kg. the two most active compounds were evaluated for their antimalarial activity against chloroquin-resistant strain (RKL9) of Plasmodium falciparum. In addition, the synthesized compounds were tested for their in-vitro antileshmanial activity against Leishmania aethiopica promastigotes and amastigotes. For both antimalarial and antileishmanial activities, compounds having an N1-p-tolyl group at the first pyrazoline ring did not require bulkiness at the second pyrazoline ring nitrogen where the compound bearing an acetyl group proved to be the most active of the whole series. On the other hand, bulkiness at the N1-position of the second pyazoline ring was necessary in case of compounds carrying the p-chlorophenyl group, where the two derivatives having an N1-butanoyl and an N1-benzoyl moieties at the second pyrazoline showed the best activity. Furthermore, the toxicity of the active compounds were tested and were proved to be non-toxic at 125, 250 and 500 mg/Kg. In addition, docking of the most active compound (having a p-tolyl group at the first pyrazoline-N and an acetyl moiety on the other pyrazoline-N) was performed against dihydrofolate reductase enzyme.

Keywords: pyrazoline derivatives, in-vivo antimalarial activity, docking, dihydrofolate reductase

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Authors: I. L. Ibrahim, A. Mann, B. M. Abdullahi

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Infectious diseases are prevalent in developing countries and plant extracts are known to contained bioactive compounds that can be used in the management of these diseases. The entire plant of Chrysanthellum indicum (Linn) was air-dried and pulverized into fine powder and then percolated to give ethanol and aqueous extracts. These extracts were phytochemically screened for metabolites and evaluated antibacterial activity against some pathogenic organisms Klebsilla, pneumonia, Bacillus subtilis, and Pseudomonas aeruginosa using agar dilution method. It was found that crude extracts of C. indicum revealed the presence of saponins, tannins, alkaloids, steroidal nucleus, cardiac glycosides, and coumarin while flavonoids and anthraquinones were absent. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the active extract of C. indicum shows that the extract could be a potential source of antibacterial agents.

Keywords: antibacterial activity, Chrysanthellum indicum, infectious diseases, phytochemical screening

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Authors: Bashair Al Riyami

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Background: Acylation stimulating protein (ASP) is a small basic protein that was isolated based on its function as a potent lipogenic factor. The role of ASP in lipid metabolism has been described in numerous studies. Several association studies suggest that ASP may play a prominent role in female fat metabolism and distribution. Progesterone is established as a female lipogenic hormone, however the mechanisms by which progesterone exert its effects are not fully understood. AIM: Since ASP is an established potent lipogenic factor with a known mechanism of action, in this study we aim to investigate acute effects of different hormone treatments on ASP levels in vivo after a fat load. Methods: This is a longitudinal study including 24 female wister rats that were randomly divided into 4 groups including controls (n=6). The rats were ovariectomized, and fourteen days later the fasting rats were injected subcutaneously with a single dose of different hormone treatments (progesterone, estrogen and testosterone). An hour later, olive was administered by oral gavage, and plasma blood samples were collected at several time points after oil administration for ASP and triglyceride measurements. Area under the curve (TG-AUC) was calculated to represent TG clearance Results: RM-ANCOVA and post-analysis showed that only the progesterone treated group had a significant postprandial ASP increase at two hours compared to basal levels and to the controls (439.8± 62.4 vs 253.45± 59.03 ug/ml), P= 0.04. Interestingly, increased postprandial ASP levels coordinated negatively with corresponding TG levels and TG-AUC across the postprandial period most apparent in the progesterone and testosterone treated groups that behaved in an opposite manner. ASP levels were 3-fold higher in the progesterone compared to the testosterone treated group, whereas TG-AUC was significantly lower in the progesterone treated group compared to the testosterone treated group. Conclusion: These findings suggest that progesterone treatment enhances ASP production and TG clearance in a simultaneous manner. The strong association of postprandial ASP levels and TG clearance in the progesterone treated group support the notion of a stimulatory role for progesterone on ASP mediated TG clearance. This is the first functional study to demonstrate a cause-effect relationship between hormone treatment and ASP levels in vivo. These findings are promising and may contribute to further understanding the mechanism of progesterone function as a female lipogenic hormone through enhancing ASP production and plasma levels.

Keywords: ASP, lipids, sex hormones, wister rats

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Authors: Khairi El Battawy, Monika Skalicki

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The present investigation has been done on pure Egyptian Arabian mares that reared in private horse studs. Fifty non-pregnant mares were selected and examined to classify them as either being reproductively healthy or subfertile mares including clinical endometritis, early embryonic death, granulosa cell tumor, repeat breeder (post-breeding endometritis), and anoestrus mares. The purpose of the study was to assess oxidative/antioxidant biochemical metabolites, lipogram, trace elements and reproductive hormones throughout reproductive conditions in mares during regular estrous, anestrum, early pregnancy, granulose cell tumor, ovulation failure, and endometritis. Results showed intensification of the free radical-dependent process in the blood of infertile mare, especially mares with endometritis. Ultrasonography as a diagnostic tool diagnosis of endometritis in mares was an important step as it revealed much information concerning infertility problem.

Keywords: endometritis, ovulation, oxidative, mare

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Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

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Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

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Authors: Benjawan Saengwichian, Alasdair E. Charles, Philip J. Hyde

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Magnetically controlled growing rods (MCGRs) have been used to stabilise and correct spinal curvature in children to support non-invasive scoliosis adjustment. Although the encapsulated driving components are intended to be isolated from body fluid contact, in vivo corrosion was observed on these components due to sealing mechanism damage. Consequently, a corrosion circuit is created with the body fluids, resulting in malfunction of the lengthening mechanism. Particularly, the chloride ions in blood plasma or cerebrospinal fluid (CSF) may corrode the MCGR alloys, possibly resulting in metal ion release in long-term use. However, there is no data available on the corrosion resistance of spinal implant alloys in CSF. In this study, an in vitro immersion configuration was designed to simulate in vivo corrosion of 440C SS-Ti6Al4V couples. The 440C stainless steel (SS) was heat-treated to investigate the effect of tempering temperature on intergranular corrosion (IGC), while crevice and galvanic corrosion were studied by limiting the clearance of dissimilar couples. Tests were carried out in a neutral artificial cerebrospinal fluid (ACSF) and phosphate-buffered saline (PBS) under aeration and deaeration for 2 months. The composition of the passive films and metal ion release were analysed. The effect of galvanic coupling, pH, dissolved oxygen and anion species on corrosion rates and corrosion mechanisms are discussed based on quantitative and qualitative measurements. The results suggest that ACSF is more aggressive than PBS due to the combination of aggressive chlorides and sulphate anions, while phosphate in PBS acts as an inhibitor to delay corrosion. The presence of Vivianite on the SS surface in PBS lowered the corrosion rate (CR) more than 5 times for aeration and nearly 2 times for deaeration, compared with ACSF. The CR of 440C is dependent on passive film properties varied by tempering temperature and anion species. Although the CR of Ti6Al4V is insignificant, it tends to release more Ti ions in deaerated ACSF than under aeration, about 6 µg/L. It seems the crevice-like design has more effect on macroscopic corrosion than combining the dissimilar couple, whereas IGC is dominantly observed on sensitized microstructure.

Keywords: cerebrospinal fluid, crevice corrosion, intergranular corrosion, magnetically controlled growing rods

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Authors: Nicholas Fletcher, Zachary Houston, Yongmei Zhao, Christopher Howard, Kristofer Thurecht

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One promising approach to enhance nanomedicine therapeutic efficacy is to include a targeting agent, such as an antibody, to increase accumulation at the tumor site. However, the application of such targeted nanomedicines remains limited, in part due to difficulties involved with biomolecule conjugation to synthetic nanomaterials. One approach recently developed to overcome this has been to engineer bispecific antibodies (BsAbs) with dual specificity, whereby one portion binds to methoxy polyethyleneglycol (mPEG) epitopes present on synthetic nanomedicines, while the other binds to molecular disease markers of interest. In this way, noncovalent complexes of nanomedicine core, comprising a hyperbranched polymer (HBP) of primarily mPEG, decorated with targeting ligands are able to be produced by simple mixing. Further work in this area has now demonstrated such complexes targeting the breast cancer marker epidermal growth factor receptor (EGFR) to show enhanced binding to tumor cells both in vitro and in vivo. Indeed the enhanced accumulation at the tumor site resulted in improved therapeutic outcomes compared to untargeted nanomedicines and free chemotherapeutics. The current work on these BsAb-HBP conjugates focuses on further probing antibody-nanomaterial interactions and demonstrating broad applicability to a range of cancer types. Herein are reported BsAb-HBP materials targeted towards prostate-specific membrane antigen (PSMA) and study of their behavior in vivo using ⁸⁹Zr positron emission tomography (PET) in a dual-tumor prostate cancer xenograft model. In this model mice bearing both PSMA+ and PSMA- tumors allow for PET imaging to discriminate between nonspecific and targeted uptake in tumors, and better quantify the increased accumulation following BsAb conjugation. Also examined is the potential for formation of these targeted complexes in situ following injection of individual components? The aim of this approach being to avoid undesirable clearance of proteinaceous complexes upon injection limiting available therapeutic. Ultimately these results demonstrate BsAb functionalized nanomaterials as a powerful and versatile approach for producing targeted nanomedicines for a variety of cancers.

Keywords: bioengineering, cancer, nanomedicine, polymer chemistry

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Authors: Y. Landkocz, P. Gosset, A. Héliot, C. Corbière, C. Vendeville, V. Keravec, S. Billet, A. Verdin, C. Monteil, D. Préterre, J-P. Morin, F. Sichel, T. Douki, P. J. Martin

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Air Pollution produced by automobile traffic is one of the main sources of pollutants in urban atmosphere and is largely due to exhausts of the diesel engine powered vehicles. The International Agency for Research on Cancer, which is part of the World Health Organization, classified in 2012 diesel engine exhaust as carcinogenic to humans (Group 1), based on sufficient evidence that exposure is associated with an increased risk for lung cancer. Amongst the strategies aimed at limiting exhausts in order to take into consideration the health impact of automobile pollution, filtration of the emissions and use of biofuels are developed, but their toxicological impact is largely unknown. Diesel exhausts are indeed complex mixtures of toxic substances difficult to study from a toxicological point of view, due to both the necessary characterization of the pollutants, sampling difficulties, potential synergy between the compounds and the wide variety of biological effects. Here, we studied the potential indirect genotoxicity of emission of Diesel engines through on-line exposure of rats in inhalation chambers to a subchronic high but realistic dose. Following exposure to standard gasoil +/- rapeseed methyl ester either upstream or downstream of a particle filter or control treatment, rats have been sacrificed and their lungs collected. The following indirect genotoxic parameters have been measured: (i) telomerase activity and telomeres length associated with rTERT and rTERC gene expression by RT-qPCR on frozen lungs, (ii) γH2AX quantification, representing double-strand DNA breaks, by immunohistochemistry on formalin fixed-paraffin embedded (FFPE) lung samples. These preliminary results will be then associated with global cellular response analyzed by pan-genomic microarrays, monitoring of oxidative stress and the quantification of primary DNA lesions in order to identify biological markers associated with a potential pro-carcinogenic response of diesel or biodiesel, with or without filters, in a relevant system of in vivo exposition.

Keywords: diesel exhaust exposed rats, γH2AX, indirect genotoxicity, lung carcinogenicity, telomerase activity, telomeres length

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Authors: Sudhanshu Sharma

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Lisinopril, 1-[N-{(s)-I-carboxy-3 phenyl propyl}-L-proline dehydrate is a lysine analog of enalaprilat, the active metabolite of enalapril. It is long-acting, non-sulhydryl angiotensin-converting enzyme (ACE) inhibitor that is used for the treatment of hypertension and congestive heart failure in daily dosage 10-80 mg. Pharmacological activity of lisinopril has been proved in various experimental and clinical studies. Owing to its importance and widespread use, efforts have been made towards the development of simple and reliable analytical methods. As per our literature survey, lisinopril in pharmaceutical formulations has been determined by various analytical methodologies like polaragraphy, potentiometry, and spectrophotometry, but most of these analytical methods are not too suitable for the Identification of lisinopril from clinical samples because of the interferences caused by the amino acids and amino groups containing metabolites present in biological samples. This report is an attempt in the direction of developing a simple and reliable method for on plate identification and quantification of lisinopril in pharmaceutical formulations as well as from human urine samples using silica gel H layers developed with a new mobile phase comprising of micellar solutions of N-cetyl-N, N, N-trimethylammonium bromide (CTAB). Micellar solutions have found numerous practical applications in many areas of separation science. Micellar liquid chromatography (MLC) has gained immense popularity and wider applicability due to operational simplicity, cost effectiveness, relatively non-toxicity and enhanced separation efficiency, low aggressiveness. Incorporation of aqueous micellar solutions as mobile phase was pioneered by Armstrong and Terrill as they accentuated the importance of TLC where simultaneous separation of ionic or non-ionic species in a variety of matrices is required. A peculiarity of the micellar mobile phases (MMPs) is that they have no macroscopic analogues, as a result the typical separations can be easily achieved by using MMPs than aqueous organic mobile phases. Previously MMPs were successfully employed in TLC based critical separations of aromatic hydrocarbons, nucleotides, vitamin K1 and K5, o-, m- and p- aminophenol, amino acids, separation of penicillins. The human urine analysis for identification of selected drugs and their metabolites has emerged as an important investigation tool in forensic drug analysis. Among all chromatographic methods available only thin layer chromatography (TLC) enables a simple fast and effective separation of the complex mixtures present in various biological samples and is recommended as an approved testing for forensic drug analysis by federal Law. TLC proved its applicability during successful separation of bio-active amines, carbohydrates, enzymes, porphyrins, and their precursors, alkaloid and drugs from urine samples.

Keywords: lisnopril, surfactant, chromatography, micellar solutions

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Authors: Subrat Kumar Bhattamisra, Vivian Lee Yean Yan, Chin Koh Lee, Chew Hui Kuean, Yun Khoon Liew, Mayuren Candasamy

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Geraniol, an acyclic monoterpenoid is the main active constituent in the essential oils of rose and palmorosa. Antioxidant, antibacterial, anticancer and antiulcer activity of geraniol was reported by many researchers. The present investigation was designed to study in vivo antiulcer and anti-Helicobacter pylori activity of geraniol. Antiulcer and anti-H. pylori activity of geraniol was evaluated on acetic acid plus H. pylori induced ulcer in rats. Acetic acid (0.03 mL) was injected to the sub-serosal layer of the stomach through laparotomy under anaesthesia. Orogastric inoculation of H. pylori (ATCC 43504) was done twice daily for 7 days. Geraniol (15 and 30 mg/kg), vehicle and standard drugs (Amoxicillin, 50 mg/kg; clarithromycin, 25 mg/kg & omeprazole, 20 mg/kg) was administered twice daily for 14 days. Antiulcer activity of geraniol was examined by the determination of gastric ulcer index, measuring the volume of gastric juice, pH and total acidity, myeloperoxidase activity and histopathological examination. Histopathological investigation for the presence of inflammation, white blood cell infiltration, edema, the mucosal damage was studied. The presence of H. pylori was detected by placing a biopsy sample from antral part of the stomach into rapid urease test. Ulcer index in H. pylori inoculated control group was 4.13 ± 0.85 and was significantly (P < 0.05) lowered in geraniol (30 mg/kg) and reference drug treated group. Geraniol increase the pH of the gastric juice (2.18 ± 0.13 in control vs. 4.14 ± 0.57 in geraniol 30mg/kg) and lower total acidity significantly (P < 0.01) in geraniol (15 & 30 mg/kg). Myeloperoxidase (MPO) activity was measured in stomach homogenate of all the groups. H. pylori control group has significant (P < 0.05) increase in MPO activity compared to normal control group. Geraniol (30 mg/kg) was showed significant (P < 0.05) and most effective among all the groups. Histopathological examination of rat stomach was scored and the total score for H. pylori control group was 8. After geraniol (30 mg/kg) and reference drug treatment, the histopathological score was significantly decreased and it was observed to be 3.5 and 2.0 respectively. Percentage inhibition of H. pylori infection in geraniol (30 mg/kg) and reference drug were found to be 40% and 50% respectively whereas, 100% infection in H. pylori control group was observed. Geraniol exhibited significant antiulcer and anti- H. pylori activity in the rats. Thus, geraniol has the potential for the further development as an effective medication in treating H. pylori associated ulcer.

Keywords: geraniol, helicobacter pylori atcc 43504, myeloperoxidase, ulcer

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Authors: Nada Bezić, Valerija Dunkić, Rušćić Mirko

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Astragalus monspessulanus L. ssp. illyricus (Bernhardt) Chater is endemic species of Fabaceae family and belongs to hemicryptophyte. This plant grows wild in the sub-Mediterranean area. We analyzed the composition of the essential oil of stems and leaves of A. monspessulanus L. ssp. Illyricus, collected in Tijarica, near Split, Croatia. Water distilled essential oils from aerial parts of investigation plant have been analysed by GC and GC/MS using VF-5ms capillary column. The total yield of oil was 0.08%, based on dry weight of samples. Thirty-eight compounds were representing 88.5% of the total oil. This essential oil was characterized by a high concentration of cis-myrtanol (20.5%), geranyl acetate (9.6%) and phytone (6.6%). Previous research in the species A. monspessulanus have included flavoalkaloids and flavonoids composition. The present study gives additional knowledge about secondary metabolites contents on the genus Astragalus.

Keywords: essential oil, isovaleric acid, Valeriana tuberosa, geranyl acetate, phytone

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Authors: Raymond Oriebe Anyasi, Harrison Atagana

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The use of a plant in the detoxification of several toxin is been known to be enhanced by various microbial endophytes which have been reported to be contained in plants growing in any contaminated soil. Plants in their natural state are mostly colonized by endophytes which in the process forms symbiotic associations with the host plants. These benefits that the endophytes offer to the plants include amongst others to: Enhance plants growth through the production of various phytohormones; increase in the resistance of environmental stresses; produce important bioactive metabolites; help in the fixing of nitrogen in the plants organelles; help in the metal translocation and accumulation in plants; assist in the production of enzymes involves the degradation of organic contaminants. Therefore recognizing these natural processes of the microbes will enable the understanding of the effective mechanism for enhanced phytoremediation. The aim of this study was to survey the progressiveness in the study involving endophyte-assisted phytoremediation of contaminants; highlighting various pollutants, the plants used, the endophytes studied as well as the type of interaction between the plants and the microbes so as to proffer a better future prospect for the technology.

Keywords: phytoremediation, endophytes, microbes, pollution, environmental management, plants

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Authors: Basavaraj Yenagi, P. Nagaraju, C. R. Patil

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Groundnut (Arachis hypohaea L.) is called as “King of oilseeds” and one of the most important food and cash crops in Indian subcontinent. Yield and quality of oil are negatively correlated with poor or imbalanced nutrition and constant exposure to both biotic and abiotic stress factors. Variety of diseases affect groundnut plant, most of them are caused by fungi and lead to severe yield loss. Imbalanced nutrition increases the concerns of environmental deterioration which includes soil fertility. Among different microbial antagonists, Pseudomonas is common member of the plant growth promoting rhizobacteria microflora present in the rhizosphere of groundnut. These are known to produce a beneficial effect on groundnut due to their high metabolic activity leading to the production of enzymes, exopolysaccharides, secondary metabolites, and antibiotics. The ability of pseudomonas lies on their ability to produce antibiotic metabolites such as 2, 4-diacetylphloroglucinol (DAPG). DAPG can inhibit the growth of fungal pathogens namely collar rot and stem rot and also increase the availability of plant nutrients through increased solubilization and uptake of nutrients. Hence, the present study was conducted for three consecutive years (2014 to 2016) in vertisol during the rainy season to assess the efficacy of DAPG producing fluorescent pseudomonas for enhancing nutrient use efficacy, bio-control of soil-borne diseases and yield of groundnut at University of Agricultural Sciences, Dharwad farm. The experiment was laid out in an RCBD with three replications and seven treatments. The mean of three years data revealed that the effect of DAPG-producing producing fluorescent pseudomonas enhanced groundnut yield, uptake of nitrogen and phosphorus and nutrient use efficiency and also found to be effective in bio-control of collar rot and stem rot incidence leading to increase pod yield of groundnut. Higher dry pod yield of groundnut was obtained with DAPG 2(3535 kg ha-1) closely followed by DAPG 4(3492 kg ha-1), FP 98(3443 kg ha-1), DAPG 1(3414 kg ha-1), FP 86(3361 kg ha-1) and Trichoderma spp. (3380 kg ha-1) over control(3173 kg ha-1). A similar trend was obtained with other growth and yield attributing parameters. N uptake ranged from 8.21 percent to FP 86 to 17.91 percent with DAPG 2 and P uptake ranged between 5.56 percent with FP 86 to 16.67 percent with DAPG 2 over control. The first year, there was no incidence of collar rot. During the second year, the control plot recorded 2.51 percent incidence and it ranged from 0.82 percent to 1.43 percent in different DAPG-producing fluorescent pseudomonas treatments. The similar trend was noticed in the third year with lower incidence. The stem rot incidence was recorded during all the three years. Mean data indicated that the control plot recorded 2.65 percent incidence and it ranged from 0.71 percent to 1.23 percent in different DAPG-producing fluorescent pseudomonas treatments. The increase in net monetary benefits ranged from Rs.5975 ha-1 to Rs.11407 ha 1 in different treatments. Hence, as a low-cost technology, seed treatment with available DAPG-producing fluorescent pseudomonas has a beneficial effect on groundnut for enhancing groundnut yield, nutrient use efficiency and bio-control of soil-borne diseases.

Keywords: groundnut, DAPG, fluorescent pseudomonas, nutrient use efficiency, collar rot, stem rot

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Authors: Farhat Jabeen, Muhammad Asad

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The study assessed the curative potential of Moringa olifera seeds against copper oxide nanoparticles induced toxicity in Oreochromis mossambicus. In order to investigate the curative potential of M. olifera seeds, firstly we examine its chemical composition, secondary metabolites, and bioactive compounds including hydroxyl-cinnamic acids, flavanols and hydroxybenzoic acids through standard methods and high performance liquid chromatography. In current study, the potential sub-lethal toxic dose of CuO-NPs (0.12 mg/l) was investigated through pilot experiment and three non-lethal doses (low=32, medium=48 and high=96 mg/l) of M. olifera were selected on the basis of its LC50 value for O. mossambicus. The experimental fish, O. mossambicus (n=100 of approximately 20 g each) were procured from Manawan Fisheries Complex, Lahore, and acclimatized for two weeks in glass aquaria. Experiment was conducted in accordance with the guidelines of Institutional Animal Ethics Committee, Government College University Faisalabad, Pakistan. During acclimatization and experimental period, fish received the commercial fish feed at 2.5% body weight daily. In order to assess the curative effect of M. olifera against CuO NPs induced toxicity, O. mossambicus were randomly divided into five groups and were designated as control (C) without any treatment, positive control (G*) exposed to potential toxic dose of CuO-NPs at 0.12 mg/l, and three treated groups namely G1, G2, and G3 co-treated with 0.12 mg/l of CuO-NPs plus different doses of M. olifera seed extract at 32, 48, and 96 mg/l, respectively for 56 days. Fish were exposed to waterborne CuO NPs and M. olifera seed extract. CuO-NPs treatment was ceased after 28 days but the doses of M. olifera were continued for 56 days. Blood was taken after 28 and 56 days through caudal venipuncture. Liver and intestine were taken for oxidative stress and histological studies after 56 days. In M. olifera seeds, moisture contents, crude protein, lipids, carbohydrates and ash were recorded as 3.8, 37.83, 32.52, 46.12, and 7.75%, respectively on dry weight basis. Total energy was recorded as 627.36 kcal/100g. Qualitative analysis of M. olifera seeds showed the presence of terpenoids, saponins, flavonoids, alkaloids and phenolics, while its quantitative analysis showed the considerable amount of total phenolics, flavonoids, saponins, and alkaloids as 134.75, 170.15, 1.57, and 0.4 µg/mg, respectively. Analysis of bioactive compounds in M. olifera seeds showed the presence of hydroxy-cinnamic acids (6.07 µg/ml), flavanols (71.72 µg/ml), and hydroxyl benzoic acids (97.82 µg/ml). The results showed that M. oliefera seed extract at 48 and 56 mg/l was able to cure against the toxic effects of CuO-NPs. The significant changes were observed in G* and G1 for sero-hepatic enzymes, anti-oxidants and histological profile. The investigations of this study showed that M. olifera is a good curative agent against potential induced toxicity of CuO-NPs in O. mossambicus. The curative effect of M. olifera is attributed to the presence of higher amount of secondary metabolites and bioactive compounds. This study suggested the use of M. olifera to curate different ailments in fish and other organisms.

Keywords: CuO nanoparticles, curative, Moringa olifera, Oreochromis mossambicus

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Authors: Martina Šutovská, Marta Jošková, Ivana Kazimierová, Lenka Pappová, Maroš Adamkov, Soňa Fraňová

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Bronchial asthma is characterized by increased bronchoconstrictor responses to provoking agonists, airway inflammation and remodeling. All these processes involve Ca2+ influx through Ca2+-release-activated Ca2+ channels (CRAC) that are widely expressed in immune, respiratory epithelium and airway smooth muscle (ASM) cells. Our previous study pointed on possible therapeutic potency of CRAC blockers using experimental guinea pigs asthma model. Presented work analyzed complex anti-asthmatic effect of long-term administered CRAC blocker, including impact on allergic inflammation, airways hyperreactivity, and remodeling and mucociliary clearance. Ovalbumin-induced allergic inflammation of the airways according to Franova et al. was followed by 14 days lasted administration of CRAC blocker (3-fluoropyridine-4-carboxylic acid, FPCA) in the dose 1.5 mg/kg bw. For comparative purposes salbutamol, budesonide and saline were applied to control groups. The anti-inflammatory effect of FPCA was estimated by serum and bronchoalveolar lavage fluid (BALF) changes in IL-4, IL-5, IL-13 and TNF-α analyzed by Bio-Plex® assay as well as immunohistochemical staining focused on assessment of tryptase and c-Fos positivity in pulmonary samples. The in vivo airway hyperreactivity was evaluated by Pennock et al. and by organ tissue bath methods in vitro. The immunohistochemical changes in ASM actin and collagen III layer as well as mucin secretion evaluated anti-remodeling effect of FPCA. The measurement of ciliary beat frequency (CBF) in vitro using LabVIEW™ Software determined impact on mucociliary clearance. Long-term administration of FPCA to sensitized animals resulted in: i. Significant decrease in cytokine levels, tryptase and c-Fos positivity similar to budesonide effect; ii.Meaningful decrease in basal and bronchoconstrictors-induced in vivo and in vitro airway hyperreactivity comparable to salbutamol; iii. Significant inhibition of airway remodeling parameters; iv. Insignificant changes in CBF. All these findings confirmed complex anti-asthmatic effect of CRAC channels blocker and evidenced these structures as the rational target in the treatment of allergic bronchial asthma.

Keywords: allergic asthma, CRAC channels, cytokines, respiratory epithelium

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Authors: Ayesha Irum, Sadia Mumtaz, Abdul Rehman, Iffat Naz, Safia Ahmed

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The present study was conducted to evaluate the potential applicability of biological trickling filter system for the treatment of simulated textile wastewater containing reactive azo dyes with bacterial consortium under non-sterile conditions. The percentage decolorization for the treatment of wastewater containing structurally different dyes was found to be higher than 95% in all trials. The stable bacterial count of the biofilm on stone media of the trickling filter during the treatment confirmed the presence, proliferation, dominance and involvement of the added microbial consortium in the treatment of textile wastewater. Results of physicochemical parameters revealed the reduction in chemical oxygen demand (58.5-75.1%), sulphates (18.9-36.5%), and phosphates (63.6-73.0%). UV-Visible and FTIR spectroscopy confirmed decolorization of dye containing wastewater was the ultimate consequence of biodegradation. Toxicological studies revealed the nontoxic nature of degradative metabolites.

Keywords: biodegradation, textile dyes, waste water, trickling filters

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Authors: Abeer Y. Ibrahim, Faten M. Ibrahim, Samah A. El-Newary, Saber F. Hendawy

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Objective: Appreciation of in-vitro anti-inflammatory and antioxidant characters of Vitex agnus-castus berries alcoholic extract and fractions, as well as in-vivo antitumor ability of alcoholic extract and chloroform fraction against Ehrlich ascites carcinoma is the aim of this study. Material and methods: Antioxidant properties of crude alcoholic extract of vitex berries as well as petroleum ether, chloroform, ethyl acetate and butanol fractions were evaluated, in-vitro assessments, as compared with standard materials, l-ascorbic acid (vitamin C) and butylated hydroxyl toluene(BHT). The anti-inflammatory activity was investigated in cyclooxygenase (COX)-1 and COX-2 inhibition assays. Moreover, in-vivo antitumor effect of vitex berries alcoholic and chloroform extracts were evaluated using Ehrlich ascites carcinoma model. Data were presented as mean±SE, and data were analyzed by one-way analysis of variance test. Results and conclusion: Berries crude extract showed potent antioxidant activity followed with its fractions ethyl acetate and chloroform as compared with standard (V.C and BHT). Ethyl acetate fraction showed good reduction capability, metal ion chelation, hydrogen peroxide scavenging, nitric oxide scavenging and superoxide anion scavenging. Meanwhile, chloroform fraction produced the highest free radical scavenging activity and total antioxidant capacity. In respectable of lipid peroxidation inhibition, crude alcoholic extract and its fractions cleared weak inhibition in comparing with standard materials. Anti-inflammatory activity of V. agnus-castus berries chloroform fraction of vitex was best COX-2 inhibitor (IC₅₀, 135.41 µg/ ml) as compared to vitex alcoholic extract or ethyl acetate fraction with weak inhibitory effect on COX-1 (IC50, 778.432 µg/ ml), where the lowest effect on COX-1 was recorded with alcoholic extract. Alcoholic extract and its fractions showed weak COX-1 inhibition activity, whereas COX-2 was inhibited (100%), compared with celecoxib drug (72% at 1000ppm). The crude alcoholic and chloroform extracts of V. agnus-castus barries significantly reduced the viable Ehrlich cell count and increased nonviable count with amelioration of all hematological parameters. This amelioration was reflected on increasing median survival time and significant increase (P < 0.05) in lifespan.

Keywords: anti-inflammatory, antioxidants, ehrlich ascites carcinoma, Vitex agnus-castus

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Authors: Stephanie Newman

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Niemann-Pick Type C disease is a rare, usually fatal lysosomal storage disorder. Although clinically characterized by progressive neurodegeneration, there is also evidence of altered innate immune responses such as neuroinflammation that promote disease progression. We have initiated an investigation into whether phagocytosis, an important innate immune activity and the process by which particles are ingested is defective in NPC. Using an in vitro assay, we have shown that NPC macrophages have a deficiency in the phagocytosis of different particles. We plan to investigate the mechanistic basis for impaired phagocytosis, the contribution that this deficiency makes to disease pathology, and whether therapies that have shown in vivo benefit are able to restore phagocytic activity.

Keywords: Niemann Pick Disease C, phagocytosis, innate immunity, lysosomal storage disorder

Procedia PDF Downloads 366

Authors: B. M. Vishal, Monamie Basu, Gopinath M., Rose Havilah Pulla

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Though there are several methods of synthesizing silver nano particles, Green synthesis always has its own dignity. Ranging from the cost-effectiveness to the ease of synthesis, the process is simplified in the best possible way and is one of the most explored topics. This study of extracting secondary metabolites from Perilla frutescens and using them for therapeutic additives has its own significance. Unlike the other researches that have been done so far, this study aims to synthesize Silver nano particles from Perilla frutescens using three available forms of the plant: leaves, seed, and commercial leaf extract powder. Perilla frutescens, commonly known as 'Beefsteak Plant', is a perennial plant and belongs to the mint family. The plant has two varieties classed within itself. They are frutescens crispa and frutescens frutescens. The species, frutescens crispa (commonly known as 'Shisho' in Japanese), is generally used for edible purposes. Its leaves occur in two forms, varying on the colors. It is found in two different colors of red with purple streaks and green with crinkly pattern on it. This species is aromatic due to the presence of two major compounds: polyphenols and perillaldehyde. The red (purple streak) variety of this plant is due to the presence of a pigment, Perilla anthocyanin. The species, frutescens frutescens (commonly known as 'Egoma' in Japanese), is the main source for perilla oil. This species is also aromatic, but in this case, the major compound which gives the aroma is Perilla ketone or egoma ketone. Shisho grows short as compared with Wild Sesame and both produce seeds. The seeds of Wild Sesame are large and soft whereas that of Shisho is small and hard. The seeds have a large proportion of lipids, ranging about 38-45 percent. Excluding those, the seeds have a large quantity of Omega-3 fatty acids, linoleic acid, and an Omega-6 fatty acid. Other than these, Perilla leaf extract has gold and silver nano particles in it. The yield comparison in all the cases have been done, and the process’ optimal conditions were modified, keeping in mind the efficiencies. The characterization of secondary metabolites includes GC-MS and FTIR which can be used to identify the components of purpose that actually helps in synthesizing silver nano particles. The analysis of silver was done through a series of characterization tests that include XRD, UV-Vis, EDAX, and SEM. After the synthesis, for being used as therapeutic additives, the toxin analysis was done, and the results were tabulated. The synthesis of silver nano particles was done in a series of multiple cycles of extraction from leaves, seeds and commercially purchased leaf extract. The yield and efficiency comparison were done to bring out the best and the cheapest possible way of synthesizing silver nano particles using Perilla frutescens. The synthesized nano particles can be used in therapeutic drugs, which has a wide range of application from burn treatment to cancer treatment. This will, in turn, replace the traditional processes of synthesizing nano particles, as this method will prove effective in terms of cost and the environmental implications.

Keywords: nanoparticles, green synthesis, Perilla frutescens, characterisation, toxin analysis

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Simakov

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In vitro organoid cultivation requires the simultaneous provision of necessary vascularization and nutrients perfusion of cells during organoid development. However, many aspects of this problem are still unsolved. The functionality of vascular network intergrowth is limited during early stages of organoid development since a function of the vascular network initiated on final stages of in vitro organoid cultivation. Therefore, a microchannel network should be created in early stages of organoid cultivation in hydrogel matrix aimed to conduct and maintain minimally required the level of nutrients perfusion for all cells in the expanding organoid. The network configuration should be designed properly in order to exclude hypoxic and necrotic zones in expanding organoid at all stages of its cultivation. In vitro vascularization is currently the main issue within the field of tissue engineering. As perfusion and oxygen transport have direct effects on cell viability and differentiation, researchers are currently limited only to tissues of few millimeters in thickness. These limitations are imposed by mass transfer and are defined by the balance between the metabolic demand of the cellular components in the system and the size of the scaffold. Current approaches include growth factor delivery, channeled scaffolds, perfusion bioreactors, microfluidics, cell co-cultures, cell functionalization, modular assembly, and in vivo systems. These approaches may improve cell viability or generate capillary-like structures within a tissue construct. Thus, there is a fundamental disconnect between defining the metabolic needs of tissue through quantitative measurements of oxygen and nutrient diffusion and the potential ease of integration into host vasculature for future in vivo implantation. A model is proposed for growth prognosis of the organoid perfusion based on joint simulations of general nutrient diffusion, nutrient diffusion to the hydrogel matrix through the contact surfaces and microchannels walls, nutrient consumption by the cells of expanding organoid, including biomatrix contraction during tissue development, which is associated with changed consumption rate of growing organoid cells. The model allows computing effective microchannel network design giving minimally required the level of nutrients concentration in all parts of growing organoid. It can be used for preliminary planning of microchannel network design and simulations of nutrients supply rate depending on the stage of organoid development.

Keywords: 3D model, consumption model, diffusion, spheroid, tissue organoid

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Authors: Valerija Dunkić, Nada Bezić

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Salvia is a genus of the well-known medicinal plant of Lamiaceae family and growing wild throughout the world. This abstract reports the comparation of the essential oils from leaves and flowers composition of Salvia pratensis L. from mountain Velebit, Croatia. Water distilled essential oils from aerial parts of investigation plant have been analysed by GC and GC/MS using VF-5ms capillary column. Fifty-three constituents, representing 99.4% of the leaf oil composition; 51 constituents, representing 86.8% of the flower oil composition. Essential oil yield varied from 0.9% to 1.3% in the leaf and flower parts of the plant. The flower essential oil was characterized by a high concentration of E-caryophyllene (21.9%) and germacrene D (10.2%). Major constituents of the leaf oil were linalool (17.7%), linalool acetate (15.3%) and limonene (9.8%). The comparative results clearly indicated that the leaf and flower oil compositions of S. pratensis were quite different in terms of major components content. The present study gives additional knowledge about secondary metabolites contents on the genus Salvia.

Keywords: essential oil, leaf, flower, Salvia pratensis L.

Procedia PDF Downloads 273

Authors: Hamza Assiry, Gamal A. Mohamed, Sabrin R. M. Ibrahim, Hossam M. Abdallah

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Chemical investigation of the aerial parts of Barleria trispinosa(Forssk.) Vahl. resulted in isolation of four major iridoids that were identified as 6,8-O,O-diacetylshanhiside methyl ester (acetyl barlerin) (1), 8-O-acetylshanzhiside methyl ester (barlerin) (2), shanzhiside methyl ester (3), and 6- ⍺ -L-rhamnopyranosyl-8-O-acetylshanzihiside methyl ester (4). The isolated compounds were confirmed by detailed one and two-dimensional NMR. Isolated compounds were tested for their cytotoxic activity on breast cancer (MCF-7, MDA-MB-231) and colon cancer (LS174T) cell linesusing sulphorhodamine B (SRB) assay. It is noteworthy that compound 1 demonstrated a significant cytotoxic potential towards MDA-MB-231 cell line with IC5016.7 ± 2.7µg / mL compared to doxorubicin whereas compounds 2, showed moderate cytotoxic potential with IC5021.2 ± 1.9µg / mL on MCF-7. The other compounds showed moderate activity on the tested cell lines.

Keywords: acanthaceae, cytotoxicity, metabolites, barleria trispinosa

Procedia PDF Downloads 124

Authors: Imam Taufik, Hilkatul Ilmi, Puryani, Mochammad Yuwono, Aty Widyawaruyanti

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Artocarpus champeden Spreng stembark (Moraceae) in Indonesia well known as ‘cempedak’ had been traditionally used for malarial remedies. The difference of growth locations could cause the difference of metabolite profiling. As a consequence, there were difference antimalarial activities in spite of the same plants. The aim of this research was to obtain the profile of metabolites that contained in A. champeden stembark from different locations in Indonesia for authentication and quality control purpose of this extract. The profiling had been performed by HPTLC-Densitometry technique and antimalarial activity had been also determined by HRP2-ELISA technique. The correlation between metabolite fingerprinting and antimalarial activity had been analyzed by Principle Component Analysis, Hierarchical Clustering Analysis and Partial Least Square. As a result, there is correlation between the difference metabolite fingerprinting and antimalarial activity from several different growth locations.

Keywords: antimalarial, artocarpus champeden spreng, metabolite fingerprinting, multivariate analysis

Procedia PDF Downloads 283

Authors: Farzad Alaeimoghadam, Farnaz Alaeimoghadam

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Plants are always being threatened by biotic and abiotic elements in their abode. Although they have inherited mechanisms to defend themselves, sometimes due to overpowering of their enemies or weakening of themselves, they just suffer from those elements. Human, as to help plants defend themselves, have developed several methods among which application of terpenes via plant biotechnology is promising. Terpenes are the most frequent and diverse secondary metabolites in plants. In these plants, terpenes are involved in different protective aspects. In this field, by utilizing biotechnological approaches on them, a delicate, precise, and an economic intervention will be achieved. In this review, first, the importance of terpenes as guardians in plants, which include their allelopathy effect, a call for alliances, and a mitigation impact on abiotic stresses will be pointed out. Second, problems concerning terpenes application in plant biotechnology comprising: damage to cell, undesirable terpene production and undesirable concentration and proportion of terpenes will be discussed. At the end, the approaches in plant biotechnology of terpenes including tampering with terpene gene sequences, compartmentalization, and localization and utilization of membrane transporters will be expressed. It is concluded with some useful notions concerning the topic.

Keywords: plant biotechnology, plant protection, terpenes, terpenoids

Procedia PDF Downloads 326

Authors: Anuranjani Kumar, Madhu Bala

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Ionising radiation exposure induces generation of free radicals and the oxidative DNA damage. SBL-1, a radioprotective leaf extract prepared from leaves Hippophae rhamnoides L. (Common name; Seabuckthorn), showed > 90% survival in mice population that was treated with lethal dose (10 Gy) of ⁶⁰Co gamma irradiation. In this study, early effects of pre-treatment with or without SBL-1 in blood peripheral blood lymphocytes (PBMCs) were investigated by cell viability assays (trypan blue and MTT). The quantitative in vitro study of Hoescht/PI staining was performed to check the apoptosis/necrosis in PBMCs irradiated at 2 Gy with or without pretreatment of SBL-1 (at different concentrations) up to 24 and 48h. Comet assay was performed in vivo, to detect the DNA strands breaks and its repair mechanism on peripheral blood lymphocytes at lethal dose (10 Gy). For this study, male mice (wt. 28 ± 2g) were administered radioprotective dose (30mg/kg body weight) of SBL-1, 30 min prior to irradiation. Animals were sacrificed at 24h and 48h. Blood was drawn through cardiac puncture, and blood lymphocytes were separated using histopaque column. Both neutral and alkaline comet assay were performed using standardized technique. In irradiated animals, alkaline comet assay revealed single strand breaks (SSBs) that showed significant (p < 0.05) increase in percent DNA in tail and Olive tail moment (OTM) at 24 h while at 48h the percent DNA in tail further increased significantly (p < 0.02). The double strands breaks (DSBs) increased significantly (p < 0.01) at 48 h in neutral assay, in comparison to untreated control. The animals pre-treated with SBL-1 before irradiation showed significantly (p < 0.05) less DSBs at 48 h treatment in comparison to irradiated group of animals. The SBL-1 alone treated group itself showed no toxicity. The antioxidant potential of SBL-1 were also investigated by in vitro biochemical assays such as DPPH (p < 0.05), ABTS, reducing ability (p < 0.09), hydroxyl radical scavenging (p < 0.05), ferric reducing antioxidant power (FRAP), superoxide radical scavenging activity (p < 0.05), hydrogen peroxide scavenging activity (p < 0.05) etc. SBL-1 showed strong free radical scavenging power that plays important role in the studies of radiation-induced injuries. The SBL-1 treated PBMCs showed significant (p < 0.02) viability in trypan blue assay at 24-hour incubation.

Keywords: radiation, SBL-1, SSBs, DSBs, FRAP, PBMCs

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Authors: Nahid H. Hajrah, Jamal S. M. Sabir, Neil Hall

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The jasmonic pathway is ubiquitous in plants and is crucial to plant development. It Is involved in fertility, ripening, and sex determination as well as in response to environmental stresses such as herbivory, pathogen drought or temperature shock. Essentially the jasmonic pathway acts to shut down growth in order to induce defence pathways. These pathways include the production of secondary metabolites which have evolved to defend against herbivores and pathogens but are of increasing interest due to their roll in medicine and biotechnology. Here we describe the transcriptional response of Rhazia stricta (a poisonous shrub widely used in traditional medicine) to jasmonic acid, in order to better characterize the genes involved in secondary metabolite production and its response to stress. We observe coordinated upregulation of flavonoid biosynthesis pathway leading to flavonols, flavones and anthocyanins but no similar coordination of the monoterpene indole alkaloid pathway.

Keywords: medicinal plants, Rhazia stricta, jasmonic acid, transcriptional analysis

Procedia PDF Downloads 115